Characterization of New Bacillus thuringiensis Isolated with Bioactivities to Tobacco Cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

담배거세미나방에 살충효과를 나타내는 새로운 Bacillus thuringiensis 균주의 특성

  • Kim, Da-A (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kim, Jin-Su (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kil, Mi-Ra (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Paek, Seung-Kyoung (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Choi, Su-Yeon (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Jin, Da-Yong (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Youn, Young-Nam (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Hwang, In-Cheon (Central Research Institute, Kyung Nong Co.) ;
  • Yu, Yong-Man (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
  • 김다아 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김진수 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 길미라 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 백승경 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 최수연 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김대용 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 윤용남 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 황인천 (㈜경농 중앙연구소) ;
  • 유용만 (충남대학교 농업생명과학대학 응용생물학과)
  • Published : 2008.03.30


Bacillus thuringiensis with selected high toxicities against tobacco cutworm, Spodoptera litura were isolated from domestic soils. When being observed under a phase-contrast microscope, the insecticidal crystal proteins were showed a bipyramidal crystal types. New CAB 109 isolate was identified to B. thuringiensis subsp. aizawai in the H serotype. As a results of insecticidal activities between CAB 109 isolate and 3 existing ready-made products against 3rd larva of S. litura, CAB 109 isolate showed 100% mortality with spore concentration $(1.3{\times}10^7cfu/ml)$. It was a very high insecticidal activity compared with a existing ready-made B. t. products. $LD_{50}$ values of CAB 109 isolate was $9.78{\times}10^5,\;6.87{\times}10^6\;and\;1.83{\times}10^7cfu/ml$ spore concentration against 2nd, 3rd and 4th larva of S. litura, respectively. Unlike Plutella xylostella, S. litura was slowly died after application up to 7 days. The weight of S. litura larva applied with CAB 109 isolate were 6-7 times less than controlled group. Even though it didn't die, it did not grow into next larva. The result observed with scanning electron microscope was that CAB 109 isolate of B. t. aizawai formed a typical bipyramidal crystal protein type. Otherwise, when CAB 109 isolate was examined with SDS-PAGE and with trypsin, there was no difference between CAB 109 strain and ready-made products of B. thuringiensis.


  1. Aronson, A.I., W. Beckman and P. Dunn. 1986. Bacillus thuringiensis and related insect pathogens. Microbil. Rev. 50: 1-24
  2. Bae, S.D., H.J., Kim, G.H., Lee and S.T., Park. 2007. Seasonal occurrence of tobacco cutworm, Spodoptera litura Fabricius and beet armyworm, Spodoptera exigua hubner using sex pheromone traps at different locations and regions in Yeongnam district. Korean J. Appl. Entomol. 46(1): 27-35
  3. Choi, J.Y., H.S. Kim, B.R. Jin, K.Y. Seol, H.Y. Park and S.K. Kang. 1996. Pathogenicity and production of Spodoptera exigua nuclear polyhedrosis virus. Korean J. Appl. Entomol. 35(3): 228-231
  4. Crickmore, N., D.R. Zeigler, J. Feitelson, E. Schnepf, J. Van rie, D. Lereclus, J. Baum and D.H. Dean. 1998. Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol. Mol. Biol. Rev. 62(3): 807-813
  5. Cho, J.R., W.R. Song., S.Y., Hwang, H.S. Kim and J.O. Lee. 1996. Age-related susceptibilty of Spodoptera litura larvae to some insecticides. Korean J. Appl. Entomol. 35(3): 249-253
  6. Gill, S.S., E.A. Cowles and V. Francis. 1995. Identification, isolation, and cloning of a Bacillus thuringiensis CryIAc toxin binding protein from the midgut of the lepidopteran insect Heliothis virescens. J. Biol. Chem. 270: 27277-27282
  7. Bae, S.D., K.B. Park and Y.J. Oh. 1997. Effect of temperature and food source on the egg and larval development of tobacco cutworm, Spodoptera litura Fabricius. Korean J. Appl. Entomol. 36(1): 48-54
  8. Raymond, M. 1985. Presentation d'un programme d'analyse logprobit pour micro-ordinnateur. Cah. ORSTOM, Ser. Ent. Med. et Parasitol. 22: 117-121
  9. Hofte, H. and H.R. Whiteley. 1989. Insecticidal crystal proteins of Bacilllus thuringiensis. Microbiol. Rev. 53(2): 242-255
  10. Goh, H.G., S.G. Lee, B.P. Lee, K.M. Choi and J.H. Kim. 1990. Simple mass-rearing of beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), on an artificial diet. Korean J. Appl. Entomol. 29: 180-183
  11. Lee, G.H., S.D. Bae, H.J. Kim, S.T. Park and M.Y. Choi. 2006. Economic injury levels for the common cutworm, Spodoptere litura (Fabricius) (Lepidoptera: Noctuidae) on soybean. Korean J. Appl. Entomol. 45(3): 333-337
  12. Kim, S.G., J.D. Park, D.I. Kim., D.J. Im., K.C. Kim and Y.M. Yu. 2003. Effects of field application of Spodoptera litura nucleopolyhedrovirus to control S. litura in chrysanthemum. Korean J. Appl. Entomol. 42(2): 153-157
  13. Goh, H.G., J.D. Park, Y.M. Choi, K.M. Cho and I.S. Park. 1991. The host plants of beet armyworm, Spodoptera exigua (Hübner), (Lepidoptera: Noctuidae) and its occurrence. Korean J. Appl. Entomol. 30(2): 111-116
  14. Tamez-Guerra, P., A.A. Iracheta, B. Pereyra-Alferez, L.J. Galan- Wong, R. Gomez-Flores, R.S. Tamez-guerra, and C. Rodriguez- Padilla. 2004. Characterization of Mexican Bacillus thuringiensis strains toxic for lepidopteran and coleopteran larvae. J. Invertebr. Pathol. 8: 7-18
  15. Da Silva, S.M.B., J.O. Silva-Werneck, R. Falcao, A.C. Gomes, R.R. Fragoso, M.T. Quezado, O.B.O. Neto, J.B. Aguiar, M.F. G. de Sa, A. Bravo and R.G. Monnerat. 2004. Characterization of novel Brazilian Bacillus thuringiensis strains active against Spodoptera frugiperda and other insect pests. J. Appl. Ent. 128: 102-107
  16. Goldberg, L.J. and J. Margalit. 1977. A bacterial spore demonstration rapid larvicidal activity against Anopheles serengotii, Uranotaenia unguiculata, Culex univittatus, Aedes aegypti and Culex pipiens. Mosq. News. 37: 355-358
  17. Park, H.W., H.S. Kim, D.W. Lee, Y.M. Yu, B.R. Jin and S.K. Kang. 1995. Expression and synergistic effect of three types of crystal protein genes in Bacillus thuringiensis. Biochem. Biophys. Res. Commun. 214(2): 602-607
  18. Laemmli, U. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 277: 680-685
  19. Ohba, M. and K. Aizwa. 1978. Serological identification of Bacillus thuringiensis and related bacteria isolated in Japan. J. Invertebr. Pathol. 32: 303-309
  20. Zheng, S.J., B. Henken, R.A. de Maagd, A. Purwito, F.A. Krens and C. Kik. 2005. Two different Bacillus thuringiensis toxin genes confer resistance to beet armyworm (Spodoptera exigua Hubner) in transgemic Bt-shallots (Allium cepa L.). Transgemic Research, 14: 261-272
  21. Kim, D.A., J.S. Kim, M.R. Kil, Y.N. Youn, D.S. Park and Y.M. Yu. 2006. Isolation and activity of insect pathogenic Bacillus thuringiensis strain from soil. Korean J. Appl. Entomol. 45(3): 357-362
  22. Gough, J.M., D.H. Kemp, R.J. Akhurst, R.D. Pearson and K. Kongsuwan. 2005. Identification and characterization of proteins from Bacillus thuringiensis with high toxic activity against the sheep blowfly, Lucilia cuprina. J. Invertebr. Pathol. 90: 39-46
  23. Yasutake, K., N.D. Binh, K. Kagoshima, A. Uemori, A. Ohgushi, M. Maeda, E. Mizuki, Y.M. Yu and M. Ohba. 2006. Occurrence of parasporin-producing Bacillus thuringiensis in Vietnam. Can. J. Microbiol. 52: 365-372
  24. Ahn, S.B., I.S. Kim, W.S. Cho, M.H. Lee and K.M. Choi. 1989. The Occurance of the crop insect pests from Korea in 1988. Korean J. Appl. Entomol. 28(4): 246-253
  25. Apaydin, O., A.F. Yenidunya, S. Harsa and H. Gunes. 2005. Isolation and characterization of Bacillus thuringiensis strains from different grain habitats in Turkey. World J. Microbiol. Biotechnol. 21: 285-292
  26. Finney, D.J. 1971. Probit analysis, estimation of the median effective dose. Cambridge University Press. London. 19-47
  27. Bae, S.D., B.R. Choi, Y.H. Song and H.J. Kim. 2003. Insecticide susceptibility in the different larva of tobacco cutworm, Spodoptera litura Fabricius (Lepidoptera: Noctuidae) collected in the soybean fields of Milyang, Korea. Korean J. Appl. Entomol. 42(3): 225-231

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